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bioRxiv · 10.1101/2025.03.07.642044

Integrating computational and experimental biophysics reveals novel insights into the RAD51-BRC4 interaction

Bresciani, V.·Rinaldi, F.·Franco, P.·Girotto, S.·Cavalli, A.·Langer, J. D.·Masetti, M.·Bernetti, M.
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Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=63 SRC="FIGDIR/small/642044v1_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@32e13org.highwire.dtl.DTLVardef@c60a6forg.highwire.dtl.DTLVardef@1d6fdceorg.highwire.dtl.DTLVardef@1b10541_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGRAPHICAL ABSTRACTC_FLOATNO C_FIG The interaction between the RAD51 and BRCA2 proteins is central for homologous recombination, a crucial pathway ensuring high-fidelity DNA repair. Recruitment of RAD51 involves eight highly conserved regions on BRCA2, named BRC repeats. To date, only the interaction between the fourth BRC repeat (BRC4) and the RAD51 C-terminal domain has been structurally characterized, while the complex of full-length RAD51 with the peptide still remains elusive. Here, we report an integrative experimental and in silico approach to reconstruct the conformational ensemble in solution for full length RAD51 in complex with BRC4. We combined AlphaFold2, crosslinking mass spectrometry (XL-MS) and small angle x-ray scattering (SAXS) data with molecular dynamics simulations (MD). These data show that the full-length RAD51-BRC4 complex is a mixture of compact and elongated conformations. Detailed analysis of the reweighted ensemble, achieved through the maximum entropy principle, identifies key residues at the N-terminal-BRC4 interface mediating complex conformational dynamics. Our evidence provides robust atomic-level insights into the interaction of RAD51 and BRC4. These findings are crucial for understanding the molecular features underlying the recognition between RAD51 and BRCA2, which are essential for developing therapeutic intervention strategies in cancer treatment.

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Bresciani, V., Rinaldi, F., Franco, P., Girotto, S., Cavalli, A., Langer, J. D., Masetti, M., Bernetti, M.. 2025-03-14. Integrating computational and experimental biophysics reveals novel insights into the RAD51-BRC4 interaction. https://doi.org/10.1101/2025.03.07.642044

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